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Kanokphan Panyaporn. Development of genome editing using CRISPR/Cas system in Chlamydomonas reinhardtii and scenedesmus spp.. Master's Degree(Genetics). Kasetsart University. Office of the University Library. : Kasetsart University, 2017.
Development of genome editing using CRISPR/Cas system in Chlamydomonas reinhardtii and scenedesmus spp.
Name: Kanokphan Panyaporn
Classification :.LCCS: QK569.C486
LCSH:
Chlamydomonas
LCSH:
Genomes
LCSH:
Genomics
LCSH:
Scenedesmaceae
Abstract:
Since the discovery of a new breakthrough method for genome editing, the CRISPR/Cas system using Cas9 endonuclease combining with single chimeric RNA known as guide RNA (gRNA) to generate double-stranded breaks and DNA repairs at desire locations, this system has been widely used with high efficiency in various organisms including bacteria, fungi, insects, plants and mammals. However, this system appeared to be problematic and have low efficient in microalgae. In Chlamydomonas, the expression of Cas9 protein is toxic to the algal cell, and genome editing could only be obtained through transient expression or a direct Cas9 ribonucleoprotein transfection. An alternative method for efficient genome editing in microalgae is needed. In this work, the toxicity of Cas9 protein in Chlamydomonas and Scenedesmus sp. was investigated to mitigate this problem. In vitro RNA-Cas9 binding assays showed that Cas9 protein bound to ~100 base RNA and this could potentially be the cause of the cell death. Cell toxicity was confirmed in Chlamydomonas as no transformant was obtained after many rounds of Cas9 transformation. A small number of Scenedesmus transformants were obtained with very low transformation rate, and, after molecular analysis only truncated versions of the transgene were identified indicating the Cas9 toxicity in Scenedesmus. Alternative forms of Cas9 including splitCas9 and CRISPR/Cpf1 were tested by expressing in Scenedesmus sp. Transformants expressing head-split-Cas9 and LbCpf1 and AsCpf1 were obtained. Molecular analysis showed that the transformants contained the full length of the transgenes and potentially expressed the transgenes without cell death. Further analysis is needed to confirm the protein expression their applicability for genome editing in microalgae.
Kasetsart University. Office of the University Library
Address:
Bangkok
Email:
tdckulib@ku.ac.th
Role:
Thesis Advisor
Name: Anongpat Suttangkakul
Role:
Thesis Co-Advisor
Created:
2017
Modified:
2023-06-19
Issued:
2023-06-19
วิทยานิพนธ์/Thesis
application/pdf
URL: https://kasetsart.idm.oclc.org/login?url=https://www.lib.ku.ac.th/KUthesis/2560/kanokphan-pan-all.pdf
CallNumber:
QK569.C486.K36
eng
DegreeName: Master of Science
Level: Master's Degree
Descipline: Genetics
Grantor: Kasetsart University
©copyrights Kasetsart University
RightsAccess:
ใช้เวลา
0.026972 วินาที
Kanokphan Panyaporn
| Title | Contributor | Type |
|---|---|---|
| Development of genome editing using CRISPR/Cas system in Chlamydomonas reinhardtii and scenedesmus spp.
มหาวิทยาลัยเกษตรศาสตร์ Kanokphan Panyaporn | Supachai Vuttipongchaikij Anongpat Suttangkakul | วิทยานิพนธ์/Thesis |
Supachai Vuttipongchaikij
Anongpat Suttangkakul
| Title | Creator | Type and Date Create |
|---|---|---|
| Development of genome editing using CRISPR/Cas system in Chlamydomonas reinhardtii and scenedesmus spp.
มหาวิทยาลัยเกษตรศาสตร์ Supachai Vuttipongchaikij;Anongpat Suttangkakul | Kanokphan Panyaporn | วิทยานิพนธ์/Thesis |
| Improvement of homology directed repair system in chlamydomonas reinhardtii by genetic engineering
มหาวิทยาลัยเกษตรศาสตร์ Supachai Vuttipongchaikij ;Anchalee Sirikhachornkit;Anongpat Suttangkakul | Suchada Srifa | วิทยานิพนธ์/Thesis |
| Genetic analysis of 3D root crown traits and root pulp characteristics in a partially inbred cassava population using genome-wide association studies
มหาวิทยาลัยเกษตรศาสตร์ Supachai Vuttipongchaikij;Anongpat Suttangkakul;Piya Kittipadaku | Pongsakorn Sunvittayakul | วิทยานิพนธ์/Thesis |
| Functional characterization of Domain of Unknown Function (DUF) families involved in the plant cell wall
มหาวิทยาลัยเกษตรศาสตร์ Supachai Vuttipongchaikij;Anongpat Suttangkakul | Pitchaporn Wannitikul | วิทยานิพนธ์/Thesis |